Minimal Nonspecific Amplification

Primer-dimer formation can be a significant issue for SYBR® Green-based qPCR. SYBR® Green Dye fluorescences when bound to any double-stranded DNA, including primer-dimers. If primer-dimers are present in your real-time PCR reaction, they can produce significant background fluorescence, confusing results. The Power SYBR® Green RNA-to-CT™ 1-Step Kit is designed to reduce primer-dimer formation and nonspecific amplification. Figure 1 shows the results when no-template control samples (NTC) are amplified using a set of 91 PCR primer pairs with the Power SYBR® Green RNA-to-CT™ 1-Step Kit, or with three other commercially available one-step qRT-PCR kits. The results show significant nonspecific signal in reactions that used other vendors’ kits whereas results from the reaction using the Power SYBR® Green RNA-to-CT™ 1-Step Kit shows low background signals.

Figure 1: Dissociation curves of the NTC from a 91-Assay Panel using the Power SYBR® Green RNA-to-Ct™ 1-Step Kit, as compared to the One-Step RT-PCR kits from three commercially available vendors.

High Sensitivity and Confidence

The kit can detect GAPDH gene product in as little as 0.1 pg of total RNA. Coupled with its ability to minimize nonspecific amplification, the Power SYBR® Green RNA-to-CT™ 1-Step Kit gives you confidence in data quality.